CN114272195B - Composition for resisting ultraviolet and blue light damage, and preparation method and application thereof - Google Patents
Composition for resisting ultraviolet and blue light damage, and preparation method and application thereof Download PDFInfo
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- CN114272195B CN114272195B CN202111638797.4A CN202111638797A CN114272195B CN 114272195 B CN114272195 B CN 114272195B CN 202111638797 A CN202111638797 A CN 202111638797A CN 114272195 B CN114272195 B CN 114272195B
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- 210000001215 vagina Anatomy 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
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- 235000019156 vitamin B Nutrition 0.000 description 1
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- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 1
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Abstract
The invention belongs to the technical field of cosmetics, and discloses an ultraviolet ray damage resistant composition, a preparation method and application thereof. The ultraviolet and blue light damage resistant composition comprises an artemisia capillaris flower extract, a pseudomonas fermentation product extract and a bifidobacterium fermentation product lysate. The composition consists of an artemisia capillaris flower extract, a pseudomonas fermentation product extract and a bifidobacterium fermentation product lysate, has good synergistic effect of all the components, can enhance the cohesion of epidermis and the regeneration of skin cells, effectively protects ultraviolet rays and blue light, has mild property, is easy to absorb and has no stimulation, and particularly meets the requirements of people on safety, effectiveness and zero burden of cosmetics, can be widely applied to skin care cosmetics, and can meet the requirements of current beauty trend.
Description
Technical Field
The invention relates to the technical field of cosmetics, in particular to an ultraviolet ray and blue light damage resistant composition, and a preparation method and application thereof.
Background
Medium-wave Ultraviolet (UVB) can directly act on DNA to cause DNA damage by mutation, and Reactive Oxygen Species (ROS) generated by UVB and long-wave ultraviolet (UVA, 320-420 nm wavelength, also called long-wave black spot effect ultraviolet) can indirectly damage cell nuclei and mitochondrial DNA to cause abnormal cell functions or apoptosis. ROS generated by Ultraviolet (UV) radiation can oxidize and damage proteins and lipids, causing abnormalities in the corresponding functions and structures. Uv radiation also affects skin pigment metabolism: the immediate response is an increase in reactive synthesis and redistribution of melanin; the delayed response is an increase in melanocyte number and an increase in viability. Thus, prolonged UV exposure can lead to tanning, premature aging, yellowing, spotting, wrinkling and even cancerous changes.
Blue light is a light with relatively high energy between 400-500nm, the corresponding spectrum includes blue (400-450 nm) and violet (450-500 nm) portions, while short-wave blue light between 400-480nm is a light with relatively high energy, and blue light in the wavelength can increase the toxin amount in the macular region of the eye, which seriously threatens the ocular fundus health of us. Solar radiation is the primary source of blue light, two to three times as intense as ultraviolet reaching the earth's surface, being part of the visible spectrum; and blue light is commonly present in cell phones, computers, projectors, laser pens, LED displays, fluorescent lamps, LED lamps, movie screens, and the like. Optical specialists indicate that blue light has extremely high energy, can easily penetrate through skin protection, causes more free radicals to be generated by skin, and is one of the main causes of skin aging. Recently, protection of people who surf the internet for a long time from blue light has become a cosmetic trend. However, most of the compositions for photoprotection on the market are directed against the effects of ultraviolet light, and few compositions for both ultraviolet and blue light protection are available.
Disclosure of Invention
The invention aims to overcome the defects of the background technology, and provides a composition for resisting ultraviolet and blue light injury and repairing a barrier, a preparation method and application thereof.
For achieving the purpose of the invention, the ultraviolet ray and blue light damage resistant composition comprises an artemisia capillaris flower extract, a pseudomonas fermentation product extract and a bifidobacterium fermentation product lysate.
Further, in some embodiments of the invention, the extract of artemisia capillaris flowers in the composition against ultraviolet and blue light damage: pseudomonas fermentation product extract: the volume ratio of the bifidobacterium fermentation product lysate is 1-2:1-2:5-10.
Preferably, in some embodiments of the present invention, the extract of artemisia capillaris flowers in the composition against ultraviolet and blue light damage: pseudomonas fermentation product extract: the volume ratio of the bifidobacterium fermentation product lysate is 1-2:1-2:5.
preferably, in some embodiments of the present invention, the extract of artemisia capillaris flowers in the composition against ultraviolet and blue light damage: pseudomonas fermentation product extract: the volume ratio of bifidobacterium fermentation product lysate is 2:2:5.
preferably, in some embodiments of the present invention, the extract of artemisia capillaris flowers in the composition against ultraviolet and blue light damage: pseudomonas fermentation product extract: the volume ratio of the bifidobacterium fermentation product lysate is 1:2:5.
preferably, in some embodiments of the present invention, the extract of artemisia capillaris flowers in the composition against ultraviolet and blue light damage: pseudomonas fermentation product extract: the volume ratio of bifidobacterium fermentation product lysate is 2:1:5.
further, in some embodiments of the present invention, the preparation method of the artemisia capillaris flower extract comprises: pulverizing herba Artemisiae Scopariae, adding 1, 3-propylene glycol solution into the pulverized herba Artemisiae Scopariae powder, soaking in water bath, adding cellulase at 40-55deg.C, stirring, heating to 40-60deg.C, performing microwave treatment, reflux-extracting in ultrasonic water bath, and filtering to obtain herba Artemisiae Scopariae extractive solution.
Preferably, in some embodiments of the present invention, the preparation method of the artemisia capillaris flower extract comprises: pulverizing herba Artemisiae Scopariae, adding 2-5 times of 40-60% 1, 3-propylene glycol solution into pulverized pollen powder, soaking for 1-4 hr, water-bathing at 45-53deg.C, adding cellulase, stirring, heating to 51-58 deg.C, microwave treating, reflux extracting in ultrasonic water-bath for 2-5 hr, repeating for 2-5 times, and filtering to obtain herba Artemisiae Scopariae flower extractive solution.
Further, in some embodiments of the invention, the preparation method of the pseudomonas fermentation product extract is: adding deionized water, phosphate buffer solution with pH of 7-8 and glucose aqueous solution into a culture container in sequence to obtain a fermentation substrate, adding pseudoalteromonas into the fermentation substrate, culturing for 5-15 days at 20-30 ℃, and centrifuging to separate supernatant.
Preferably, in some embodiments of the present invention, the preparation method of the pseudomonas fermentation product extract is: adding 2-4 times deionized water, 0.5-2.5 times pH7-8 phosphate buffer solution with concentration of 0.25-0.45mmol/L, and 1-4 times glucose aqueous solution with concentration of 20-60wt% into a culture container in sequence to obtain fermentation substrate, adding pseudoalteromonas into the fermentation substrate, culturing at 25-37deg.C for 5-13 days, and centrifuging to separate supernatant.
Further, in some embodiments of the invention, the bifidobacterium fermentation product lysate is prepared by: adding deionized water, phosphate buffer solution with pH of 7-8 and glucose aqueous solution into a culture container in sequence to obtain a fermentation substrate, adding bifidobacterium into the fermentation substrate, and culturing for 5-15 days under the condition of room temperature and oxygen-free condition; adding N-acetylmuramidase hydrolase, stirring, standing, and separating supernatant.
Preferably, in some embodiments of the present invention, the preparation method of the bifidobacterium fermentation product lysate is: adding 2-5 times deionized water, 0.5-1.5 times pH7-8 phosphate buffer solution with concentration of 0.25-0.45mmol/L and 1-4 times glucose aqueous solution with concentration of 25-60wt% into a culture container in sequence to obtain fermentation substrate, adding bifidobacterium into the fermentation substrate, culturing for 5-13 days under anaerobic condition at 25-37 ℃, adding N-acetylmuramidase with total mass of 3-10%, stirring for 0.5-4h, standing, and separating supernatant.
Herba Artemisiae Scopariae (Artemisia capillaris thunder.) is a herb of Artemisia of Compositae, and has strong fragrance. The main root is obvious woody, the stem is single or few, the height can reach 120 cm, the reddish brown or brown, basal leaves are densely planted, and the rosettes are often formed. Herba Artemisiae Scopariae is rich in vitamin C and vitamin B, contains multiple microelements and more than 20 amino acids required by human body, and has good health promoting effect. Because it is rich in vitamins and minerals, it has strong vitality and can survive throughout winter, also called artemisia annua. The artemisia capillaris flower extract is rich in beta-carotene and various vitamins, and can eliminate active oxygen and effectively prevent skin aging.
Pseudoalteromonas, the origin is China. Rod-like, non-forming microvesicles and spores, most moving with polar flagella, is a gram-negative bacterium living in coastline as well as in the ocean. The pseudomonas fermentation product extract obtained by the method is rich in various proteoglycans and extracellular polysaccharide, can efficiently preserve moisture, promote epidermis regeneration and strengthen the integrity of skin structures.
Bifidobacterium is a gram-positive genus of bacteria, and the cells are in the shape of rods, sometimes bifurcated at one end, and strictly anaerobic, and widely exist in biological environments such as the digestive tract, vagina, and oral cavity of humans and animals. Bacteria of the genus bifidobacterium are one of the important constituent members of the intestinal flora of humans and animals and can be used as probiotics in food, medicine and feed. The bifidobacterium fermentation product lysate obtained by the invention contains metabolites, cytoplasmic components, cell wall components and polysaccharide complexes, and can specifically support the protection and repair mechanism of skin, so that the bifidobacterium fermentation product lysate can resist ultraviolet rays and damage caused by blue light.
On the other hand, the invention also provides a preparation method of the ultraviolet-resistant and blue-ray damage-resistant composition, which comprises the following steps: extracting herba Artemisiae Scopariae flower to obtain extract: pseudomonas fermentation product extract: mixing the lysate of the fermentation product of the bifidobacterium, homogenizing, adding a clarifying agent, standing, centrifugally filtering, and concentrating under reduced pressure to obtain the composition for resisting ultraviolet and blue light damage and repairing the barrier.
Further, in some embodiments of the invention, the reduced pressure concentration is 1/6 to 1/2 of the original weight; preferably, in some embodiments of the invention, the filtration is with a microfiltration membrane of 0.1-1.0 μm.
In yet another aspect, the present invention also provides the use of the aforementioned ultraviolet light damage resistant composition, said use being: the composition for preventing ultraviolet and blue light damage is used in cosmetics or cosmetic additives.
In yet another aspect, the present invention also provides a cosmetic comprising the aforementioned composition for protecting against ultraviolet and blue light damage.
Preferably, the ultraviolet and blue light damage resistant composition accounts for 0.5-5% of the cosmetic by mass.
Preferably, the cosmetic is an aqueous solution, an emulsion, a spray, a cream, a gel or a mask.
In yet another aspect, the present invention also provides a concentrate of a composition for protecting against uv and blue light damage, repairing a barrier, the concentrate comprising the aforementioned uv and blue light damage composition, water, glycerin, butylene glycol, dipropylene glycol, erythritol, bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate, p-hydroxyacetophenone, arginine, polymethylsilsesquioxane, C10-30 alkanol acrylate cross-linked polymer, panthenol, ethylhexyl glycerin, carbomer, dipotassium glycyrrhizinate, and disodium EDTA.
Blue light irradiation induces ROS to generate, so that the synthesis of type I collagen in skin cells is reduced, MMPI synthesis is increased, collagen is further degraded, and skin photoaging is induced. When the skin is irradiated by blue light, the cells are damaged to generate a large amount of ROS, and many active substances in the cells are denatured. The artemisia capillaris flower extract prepared by the invention can directly inhibit the release of inflammatory factor COX-2 generated by injury caused by ROS, reduce inflammation, resist blue light burn/sunburn cells, and reduce DNA methylation and protein carbonylation of cells caused by ultraviolet rays/blue light; the bifidobacterium lysate can inhibit the secretion of IL-10, improve the secretion of IL-12, regulate and control the DNA repair process of dermal cells, weaken the aggregation of immune cells around injured cells caused by COX-2 secretion, and strengthen the effect of the artemisia capillaris flower extract; the pseudoalteromonas fermentation product extract can activate desmosome-3, promote the generation of type I collagen and type III collagen, has a good repairing effect on dermis injury caused by active oxygen, and can obviously improve the content of bromodeoxyadenosine after being compounded with the artemisia capillaris flower extract and the bifidobacterium lysate, so that the cohesion of epidermis and regeneration of skin cells can be enhanced after the three are compounded, and the product has an excellent effect on resisting blue light injury.
Drawings
FIG. 1 is a graph showing the results of experiments on cellular DNA repair using different compositions according to examples of the present invention;
fig. 2 is a skin elasticity test result in the embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. It is to be understood that the following description is intended to be illustrative of the invention and not restrictive.
The terms "comprising," "including," "having," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, step, method, article, or apparatus.
When an equivalent, concentration, or other value or parameter is expressed as a range, preferred range, or a range bounded by a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when ranges of "1 to 5" are disclosed, the described ranges should be construed to include ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a numerical range is described herein, unless otherwise indicated, the range is intended to include its endpoints and all integers and fractions within the range.
The indefinite articles "a" and "an" preceding an element or component of the invention are not limited to the requirement (i.e. the number of occurrences) of the element or component. Thus, the use of "a" or "an" should be interpreted as including one or at least one, and the singular reference of an element or component includes the plural reference unless the amount clearly dictates otherwise.
Furthermore, the descriptions of the terms "one embodiment," "some embodiments," "examples," "particular examples," or "some examples," etc., described below mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily for the same embodiment or example. The technical features of the respective embodiments of the present invention may be combined with each other as long as they do not collide with each other.
Example 1
A composition for resisting ultraviolet and blue light injury and repairing barrier is prepared by the following steps:
(1) Pulverizing herba Artemisiae Scopariae;
(2) Adding 3 times of 50% 1, 3-propanediol solution into pulverized pollen powder of herba Artemisiae Scopariae, respectively, and soaking for 2 hr. Adding cellulase in water bath at 48deg.C, stirring for 20min, heating to 55deg.C, microwave treating for 10min, reflux extracting in ultrasonic water bath for 3 hr, repeating for 3 times, and filtering to obtain herba Artemisiae Scopariae flower extractive solution
(3) Adding 3 times deionized water, 1 time phosphate buffer solution with pH of 7-8 and concentration of 0.35mmol/L and 3 times glucose aqueous solution with concentration of 50wt% into a culture container in sequence to obtain fermentation substrate. Adding bifidobacteria into the fermentation substrate, and culturing for 10 days under the condition of room temperature and oxygen-free condition; adding N-acetylmuramyl hydrolase accounting for 5% of the total mass of the raw materials, stirring for 2 hours, and standing to separate supernatant;
(4) Adding 5 times deionized water, 1.5 times phosphate buffer solution with pH of 7-8 and concentration of 0.35mmol/L and 3 times glucose aqueous solution with concentration of 30-50wt% into a culture container in sequence to obtain fermentation substrate. Adding pseudoalteromonas to the fermentation substrate, culturing for 10 days at 25 ℃, and centrifuging to separate supernatant;
(5) Mixing the three liquids obtained in the steps (2), (3) and (4), homogenizing, adding a clarifying agent, standing, centrifuging for 20 minutes, filtering with a microfiltration membrane with the pore diameter of 0.1-1.0 μm, concentrating under reduced pressure to 1/3 of the original weight, and obtaining the composition for resisting ultraviolet and blue light damage and repairing the barrier.
Example 2
Testing of human skin fibroblasts against oxidative damage by blue light radiation by combination of artemisia capillaris flower extracts, pseudoalteromonas fermentation product extracts, bifidobacterium fermentation product lysates in different proportions: the artemisia capillaris flower extract, the pseudomonas fermentation product extract and the bifidobacterium fermentation product lysate are compounded according to the following table proportions to obtain compositions 1-12, comparative examples 1-3, wherein the compounding proportions are shown in the table 1, and the numerical values of the proportions are the artemisia capillaris flower extract: pseudomonas fermentation product extract: bifidobacterium fermentation product lysate.
Table 1 compositions 1 to 12, comparative examples 1 to 3, wherein the three components were compounded in proportions
| Composition (volume ratio) | |
| Composition 1 | 1:1:5 |
| Composition 2 | 1:2:5 |
| Composition 3 | 2:1:5 |
| Composition 4 | 2:2:5 |
| Composition 5 | 1:1:8 |
| Composition 6 | 1:2:8 |
| Composition 7 | 2:1:8 |
| Composition 8 | 2:2:8 |
| Composition 9 | 1:1:10 |
| Composition 10 | 1:2:10 |
| Composition 11 | 2:1:10 |
| Composition 12 | 2:2:10 |
| Comparative example 1 | 1:0:0 |
| Comparative example 2 | 0:1:0 |
| Comparative example 3 | 0:0:1 |
Note that: the concentration of the artemisia capillaris flower extract, the pseudoalteromonas fermentation product extract and the bifidobacterium fermentation product lysate in each group is the same, and the addition proportion is different.
In the above-described compositions of the present invention and comparative examples, the compositions were tested for oxidative damage to blue light as follows:
the invention is judged by measuring the level of active oxidative species in the composition on human fibroblasts exposed to blue light. The blue light source composed by the LED generates stresses at 415nm and 470 nm. Reactive Oxidative Species (ROS) generated by such exposure are measured at the cellular level by fluorescent probes.
(1) Normal human skin fibroblasts in culture were pretreated with 0.1% mass fraction composition 1-12, composition in comparative examples 1-3 twice daily by diluting to 0.1% volume in specific medium, while control cultures were maintained in untreated condition. After 24 hours, the control culture and a portion of the treated culture were exposed to HEV blue light (415 and 470nm, at 3mW/cm 2 For 18 minutes below) while the other part is maintained protected from light. After this exposure, compositions 1-12, compositions of comparative examples 1-3 were applied twice daily for 24 hours again. Then, the exposure to blue light is repeated again, followed by the test for ROS detection.
(2) Cells were left to stand at 37℃for 30min in the presence of fluorescent probes (CellROX green reagent, sieimer's fly). After fixation and washing, the cells were observed under an epifluorescence microscope (zeissaxioert 200M microscope). Based on the obtained photographs, quantification of the fluorescence intensity, which is proportional to the amount of ROS present, is performed by means of a Volocity image analysis software (PerkinElmer, inc.). The experimental results are shown in table 2.
Table 2 composition tests for oxidative damage to blue light
The above test results show that the compositions of test examples 1-12 can greatly reduce the effect of ROS generated by human skin fibroblasts after blue light irradiation, and have excellent blue light resisting effect. However, the composition ratio is different under the same conditions, and the reduction of ROS can be influenced, wherein the ROS reducing effect of the compositions 4, 2 and 3 is relatively good.
Example 3
The experiments of the different proportions of artemisia capillaris flower extract, pseudomonas fermentation product extract and bifidobacterium fermentation product lysate compounding on human skin explant cells for resisting ultraviolet/blue light radiation induced protein carbonylation are carried out by adopting a DNPH colorimetry method, and the results are shown in Table 3.
(1) Sample pretreatment: taking a certain amount of human skin explant cells, adding 1% of each composition sample by mass fraction, respectively placing the samples under ultraviolet light and HEV blue light irradiation for 6 hours in an experiment group, rinsing the blank group in ice physiological saline after the treatment is finished, adding a certain amount of ice HEPES buffer solution, and preparing homogenate with the mass fraction of 10%, wherein the pH value of the HEPES buffer solution is 7.4. Centrifuging the homogenate at a rotation speed of 500g for 15min, taking a supernatant, transferring the supernatant into a 10ml centrifuge tube, adding 0.1kg/L streptomycin sulfate solution into the supernatant to obtain a final concentration of 0.0L kg/L, namely, the volume ratio of the streptomycin sulfate solution to the supernatant is 1:9. after 10min of standing at room temperature, centrifuging for 10min at 11000g, and collecting supernatant.
(2) Protein carbonylation assay: to 100. Mu.l of the supernatant was added 400. Mu.l of 10mmol/L DNPH (dissolved with 2mol/L HCL). Each reaction system was placed in the dark for l h and stirred 1 time every 10 min. Mu.l of 0.2kg/L trichloroacetic acid (TCA) solution was added, centrifuged at 12000g for 15min and the supernatant was discarded. The resulting precipitate was washed 3 times with 1ml of a mixture of ethanol and ethyl acetate (V/v=1:1), and the final precipitate was dissolved with 1.25ml of 6mol/L guanidine hydrochloride (37 ℃, water bath 15 mitt). 12000g, 15rain was centrifuged, and the supernatant was collected. The absorbance was measured by the Biuret method at 370nm using an ultraviolet-visible spectrophotometer.
(3) Taking the fluorescence change rate as an experimental result, the calculation mode is as follows:
rate of change= (suction) Experimental group Suction-suction Blank group ) Suction/suction device Blank group *100％
Suction pipe Experimental group : absorbance of experimental group
Suction pipe Blank group : absorbance of blank group
TABLE 3 test results of protein carbonylation induced by UV/blue radiation
| Test item | Ultraviolet irradiation (%) | HEV blue light irradiation (%) |
| Composition 1 | 58.1 | 32.9 |
| Composition 2 | 64.9 | 35.3 |
| Composition 3 | 63.6 | 34.5 |
| Composition 4 | 66.3 | 35.7 |
| Composition 5 | 50.5 | 28.6 |
| Composition 6 | 55.6 | 31.8 |
| Composition 7 | 53.8 | 31.2 |
| Composition 8 | 59.5 | 33.8 |
| Composition 9 | 48.9 | 26.9 |
| Composition 10 | 51.3 | 30.1 |
| Composition 11 | 52.7 | 30.2 |
| Composition 12 | 57.7 | 32.4 |
| Comparative example 1 | 34.7 | 17.9 |
| Comparative example 2 | 31.8 | 15.7 |
| Comparative example 3 | 36.4 | 20.3 |
Table 3 shows that the compositions of test examples 1-12 are effective in reducing protein carbonylation due to UV and blue light, with compositions 4, 2, 3 having a relatively better reduction. Therefore, the combination of the artemisia capillaris flower extract, the pseudomonas fermentation product extract and the bifidobacterium fermentation product lysate can be judged to be capable of effectively inhibiting ultraviolet and blue light damage.
Example 4
Different proportions of the artemisia capillaris flower extract, the pseudomonas fermentation product extract and the bifidobacterium fermentation product lysate are compounded to test the human skin explant cells against ultraviolet/blue light radiation induced pro-inflammatory mediator cyclooxygenase-2 (COX-2), and the test results are shown in Table 4.
The test method is as follows: 1% mass fraction of each composition was topically applied to an HEV light exposure (5.76J/cm 2 ) Human skin explants (25 year old donor) were left on the bottom for 2 hours a day for 3 days without any composition. Cyclooxygenase-2 (COX-2) levels in human skin explant cells were measured using a double antibody sandwich method. By purified human epoxyCoating a microplate with an enzyme-2 (COX-2) antibody to prepare a solid-phase antibody, sequentially adding cyclooxygenase-2 (COX-2) into the microwells of the coated monoclonal antibody, combining with an HRP-labeled cyclooxygenase-2 (COX-2) antibody to form an antibody-antigen-enzyme-labeled antibody complex, and adding a substrate TMB for color development after thorough washing. TMB is converted to blue under the catalysis of HRP enzyme and to final yellow under the action of acid. The shade of color was positively correlated with cyclooxygenase-2 (COX-2) in the sample. The absorbance was measured at a wavelength of 450nm using an enzyme-labeled instrument, and the rate of change of absorbance in the experimental group and the blank group was used as the experimental result, calculated as follows:
rate of change= (suction) Experimental group Suction-suction Blank group ) Suction/suction device Blank group *100％
Suction pipe Experimental group : absorbance of experimental group
Suction pipe Blank group : absorbance of blank group
TABLE 4 results of the ultraviolet/blue radiation resistant induced cyclooxygenase-2 test of pro-inflammatory mediators
| Test item | HEV blue light irradiation (%) |
| Composition 1 | 27.9 |
| Composition 2 | 30.3 |
| Composition 3 | 29.8 |
| Composition 4 | 30.7 |
| Composition 5 | 23.6 |
| Composition 6 | 26.8 |
| Composition 7 | 26.2 |
| Composition 8 | 28.8 |
| Composition 9 | 21.9 |
| Composition 10 | 25.1 |
| Composition 11 | 25.2 |
| Composition 12 | 27.4 |
| Comparative example 1 | 16.8 |
| Comparative example 2 | 14.6 |
| Comparative example 3 | 19.1 |
The results shown in Table 4 demonstrate that the compositions of test examples 1-12 are effective in reducing COX-2 expression and reducing the production of inflammatory responses, with compositions 4, 2, and 3 having a relatively better attenuation. Therefore, the combination of the artemisia capillaris flower extract, the pseudomonas fermentation product extract and the bifidobacterium fermentation product lysate can be judged to be capable of effectively inhibiting ultraviolet and blue light damage.
Example 5
Experiments of different compositions for cellular DNA repair: bromodeoxyuridine (BrdU) is a thymidine analog. The amount of BrdU in DNA represents the extent of DNA repair. The composition was applied to the breast skin using a bovine breast skin model at a mass fraction of 1%, and the blank was continuously irradiated with HEV blue light for 3 hours without adding any composition, and the bromodeoxyuridine (BrdU) content was measured. The experimental results are shown in figure 1.
The experimental result of figure 1 shows that the combination of the artemisia capillaris flower extract, the pseudomonas fermentation product extract and the bifidobacterium fermentation product lysate can strengthen the repair of cell DNA damage.
Example 6
Human body efficacy test: in a specific embodiment, the invention provides a formulation process of essence of a composition for resisting ultraviolet light and blue light damage and repairing a barrier, and the specific formulation is shown in table 5.
Table 5 essential oil formulations containing the compositions of the present invention
Compositions 4, 2, and 3 were added as shown in Table 5 to prepare essence samples 1-3, respectively.
Example 7
MTT cytotoxicity test: in 96well Multiplate (burning) according to 1X10 4 The density of cells/well was inoculated with 100. Mu.L each of DMEM medium containing 10% bovine serum and keratinocytes (HaCaT), and cultured for 24 hours and then replaced with serum-free medium. The above-mentioned substances are added into serum-free culture medium respectivelyCompositions 4, 2, and 3 in the examples were cultured for 24 hours after treatment. The medium was then removed, treated with 20. Mu.L of MTT solution and allowed to react at 37℃for 2 hours. 200 mu L of isopropanol was added to the MTT solution-removed cells, gently swirled for 30min, the crystalline formazan was completely dissolved, absorbance was measured at 570nm, and cell viability was calculated according to the following formula.
The control group was tested without the addition of sample. The cytotoxicity-related results are shown in Table 6.
TABLE 6MTT cytotoxicity test results
Example 8
Safety test (human skin patch test): selecting 15 healthy subjects without skin disease allergy history between 20 and 50 years old, selecting qualified patch testers, dripping about 15 mu L of samples 1 to 3 into the patch testers in a closed patch test mode, attaching a special adhesive tape to the back of the subjects by external application, attaching 20 patch testers to each subject, respectively attaching muscle base fluid samples of the samples 1 to 3, removing the testers after 24 hours, observing skin reactions after removing the testers at 0.5, 6, 12, 24 and 48 hours, and recording the results according to skin reaction grade standards in skin care product health Specification.
Test results: the results of the human skin patch test show that all subjects pass the patch test and observe skin reactions at 0.5, 6, 12, 24 and 48 hours, wherein adverse reactions such as skin erythema, pimple and blister appear in 0 cases, which indicates that the product of the invention is safe and has no stimulation.
Example 9
Skin elasticity test: since protein carbonylation resulted in collagen fiber hardening and loss of skin tissue softness, and thus skin elasticity, the effect of the composition on blue light repair was reflected by detection of skin elasticity change after HEV blue light irradiation, and 45 30-55 year old female volunteers were selected and divided into three groups, and facial skin elasticity was measured after exposure to HEV blue light for a period of time (minimum 4 hours per day) before use of test samples 1, 2, 3 and after continuous use of cosmetics for 28 days. The test subjects used 2 times per day of essence at the selected part, cleaned the facial skin, wiped the skin, smeared the essence on the facial skin by hands, and used for four weeks continuously. The subject did not use other products during the test. The higher the variation compared with the variation before the test, the better the softness improvement of the skin is, and the average value of the data is shown in fig. 2.
From figure 2, it can be seen that the combination of herba Artemisiae Scopariae flower extract, pseudomonas fermentation product extract, and Bifidobacterium fermentation product lysate can improve skin elasticity, and relieve protein carbonylation effect of blue light radiation on human fibroblast.
Example 10
Wrinkle detection, evaluation of efficacy to improve eye wrinkles, test population: 45 people; test site: an eye; 45 volunteers (except pregnant or lactating women) aged 18-60 years and having obvious wrinkles on the eyes were selected and divided into 3 groups, and samples 1-3 were used each time in the morning and evening, and the total size of wrinkles at the corners of the eyes was measured with Antera 3D (Miravex, ireland) before, 2W after and 4W after the use of the product. The subjects need to clean the face, sit still for 20min in a laboratory with the temperature of 21+/-1 ℃ and the relative humidity of 50+/-5%, then measure the total size of the wrinkles at the corners of eyes, measure each area in parallel for 3 times, and take the average value; the efficacy of improving the eye wrinkles is evaluated by the rate of increase in the overall size of the eye wrinkles, the more the overall size of the eye wrinkles is reduced, the better the effect.
The results were averaged, and the experimental results are shown in table 7.
TABLE 7 increase and decrease in the total size of eye corner wrinkles (%)
| Sample 1 | Sample 2 | Sample 3 | |
| Before use | 0 | 0 | 0 |
| 2W | -15.9 | -15.6 | -14.7 |
| 4W | -20.4 | -18.5 | -17.8 |
It will be readily appreciated by those skilled in the art that the foregoing is merely illustrative of the present invention and is not intended to limit the invention, but any modifications, equivalents, improvements or the like which fall within the spirit and principles of the present invention are intended to be included within the scope of the present invention.
Claims (16)
1. An ultraviolet light damage resistant composition, which is characterized by comprising an artemisia capillaris flower extract, a pseudomonas fermentation product extract and a bifidobacterium fermentation product lysate; the virgate wormwood flower extract in the ultraviolet light damage resistant composition comprises the following components: pseudomonas fermentation product extract: the volume ratio of the bifidobacterium fermentation product lysate is 1-2:1-2:5-10; the preparation method of the artemisia capillaris flower extract comprises the following steps: pulverizing herba Artemisiae Scopariae, adding 1, 3-propylene glycol solution into the pulverized herba Artemisiae Scopariae powder, soaking in water bath, adding cellulase at 40-55deg.C, stirring, heating to 40-60deg.C, performing microwave treatment, reflux-extracting in ultrasonic water bath, and filtering to obtain herba Artemisiae Scopariae extract; the preparation method of the pseudoalteromonas fermentation product extract comprises the following steps: adding deionized water, phosphate buffer solution with pH of 7-8 and glucose aqueous solution into a culture container in sequence to obtain a fermentation substrate, adding pseudoalteromonas into the fermentation substrate, culturing for 5-15 days at 20-30 ℃, and centrifuging to separate supernatant; the preparation method of the bifidobacterium fermentation product lysate comprises the following steps: adding deionized water, phosphate buffer solution with pH of 7-8 and glucose aqueous solution into a culture container in sequence to obtain a fermentation substrate, adding bifidobacterium into the fermentation substrate, and culturing for 5-15 days under the condition of room temperature and oxygen-free condition; adding N-acetylmuramidase hydrolase, stirring, standing, and separating supernatant.
2. The ultraviolet and blue light damage resistant composition according to claim 1, wherein the extract of artemisia capillaris flowers in the ultraviolet and blue light damage resistant composition: pseudomonas fermentation product extract: the volume ratio of the bifidobacterium fermentation product lysate is 1-2:1-2:5.
3. the ultraviolet and blue light damage resistant composition according to claim 1, wherein the extract of artemisia capillaris flowers in the ultraviolet and blue light damage resistant composition: pseudomonas fermentation product extract: the volume ratio of bifidobacterium fermentation product lysate is 2:2:5.
4. the ultraviolet and blue light damage resistant composition according to claim 1, wherein the extract of artemisia capillaris flowers in the ultraviolet and blue light damage resistant composition: pseudomonas fermentation product extract: the volume ratio of the bifidobacterium fermentation product lysate is 1:2:5.
5. the ultraviolet and blue light damage resistant composition according to claim 1, wherein the extract of artemisia capillaris flowers in the ultraviolet and blue light damage resistant composition: pseudomonas fermentation product extract: the volume ratio of bifidobacterium fermentation product lysate is 2:1:5.
6. the composition for resisting ultraviolet and blue light damage according to claim 1, wherein the preparation method of the artemisia capillaris flower extract comprises the following steps: pulverizing herba Artemisiae Scopariae, adding 2-5 times of 40-60% 1, 3-propylene glycol solution into pulverized pollen powder, soaking for 1-4 hr, water-bathing at 45-53deg.C, adding cellulase, stirring, heating to 51-58 deg.C, microwave treating, reflux extracting in ultrasonic water-bath for 2-5 hr, repeating for 2-5 times, and filtering to obtain herba Artemisiae Scopariae flower extractive solution.
7. The ultraviolet and blue light damage resistant composition of claim 1, wherein the pseudomonas fermentation product extract is prepared by the steps of: adding 2-4 times deionized water, 0.5-2.5 times pH7-8 phosphate buffer solution with concentration of 0.25-0.45mmol/L, and 1-4 times glucose aqueous solution with concentration of 20-60wt% into a culture container in sequence to obtain fermentation substrate, adding pseudoalteromonas into the fermentation substrate, culturing for 5-13 days at 25-30deg.C, and centrifuging to separate supernatant.
8. The composition for resisting ultraviolet and blue light damage according to claim 1, wherein the preparation method of the bifidobacterium fermentation product lysate comprises the following steps: adding 2-5 times deionized water, 0.5-1.5 times pH7-8 phosphate buffer solution with concentration of 0.25-0.45mmol/L and 1-4 times glucose aqueous solution with concentration of 25-60wt% into a culture container in sequence to obtain fermentation substrate, adding bifidobacterium into the fermentation substrate, culturing for 5-13 days under anaerobic condition at room temperature, adding N-acetylmuramidase with total mass of 3-10%, stirring for 0.5-4h, standing, and separating supernatant.
9. A process for the preparation of a composition for protection against uv and blue light damage according to any one of claims 1 to 8, characterized in that it comprises: mixing herba Artemisiae Scopariae flower extract, pseudomonas fermentation product extract, and Bifidobacterium fermentation product lysate, homogenizing, adding clarifier, standing, centrifuging, filtering, and concentrating under reduced pressure to obtain ultraviolet and blue light injury resisting composition.
10. The method of preparing a composition for preventing ultraviolet and blue light damage according to claim 9, wherein the concentration under reduced pressure is 1/6 to 1/2 of the original weight.
11. A method of preparing a composition for protection against uv and blue light damage according to claim 9, wherein said filtration is with a microfiltration membrane of 0.1-1.0 μm.
12. Use of the composition for protection against uv and blue light damage according to any one of claims 1 to 8, characterized in that it is: use of the composition according to any one of claims 1 to 8 for the preparation of a cosmetic or cosmetic additive.
13. A cosmetic product comprising the composition of anti-uv and blue light damage according to any one of claims 1 to 8.
14. The cosmetic according to claim 13, wherein the composition for preventing ultraviolet and blue light damage comprises 0.5-5% by mass of the cosmetic.
15. The cosmetic product according to claim 13, wherein the cosmetic product is an aqueous solution, an emulsion, a spray, a cream, a gel or a mask.
16. An anti-ultraviolet and blue light damage essence, comprising the anti-ultraviolet and blue light damage composition of any one of claims 1-8, water, glycerin, butylene glycol, dipropylene glycol, erythritol, bis-diethoxydiglycol cyclohexane 1, 4-dicarboxylate, p-hydroxyacetophenone, arginine, polymethylsilsesquioxane, C10-30 alkanol acrylate cross-linked polymer, panthenol, ethylhexyl glycerin, carbomer, dipotassium glycyrrhizinate, and disodium EDTA.
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